The Barka desalination plant, commissioned in 2018, is the largest desalination plant in Oman. It has a capacity of 281 MLD with a reverse osmosis (RO) first-pass recovery rate of 46%. As part of the standard operator practice, a membrane autopsy was conducted to determine the cause of reductions in membrane performance. This study investigated fouled membranes (model No. SW30HRLE-440) from two different locations in the membrane rack. Various analytical methods were used to conduct the membrane autopsy. Field-emission scanning electron microscopy/energy-dispersive X-ray (FESEM/EDS) analyses of membrane samples showed major components of inorganic foulants. Moreover, black and salt-like crystals deposited on the membrane surface revealed significant carbon (C) components and oxygen (O), with a small amount of magnesium (Mg), chloride (Cl), sodium (Na), aluminium (Al), and calcium (Ca), respectively. A Fourier transform infrared (FTIR) analysis revealed the presence of long-chain hydrocarbons, carboxylic acids/esters, carbohydrates/polysaccharides, and inorganic foulants. Thermogravimetric analyses (TGA) of the membranes showed a high initial weight loss due to organic and inorganic fouling. X-ray photoelectron (XPS) analyses further confirmed the presence of inorganic and organic foulants on the membrane surfaces. Bacteria identification results showed the presence of Bacillus cereus and Bacillus marisflavi. This paper offers a detailed analysis of the foulants present on the reverse osmosis membrane surface and sub-surface before and after a cleaning process.